Carmo Helena, Brulport Marc, Hermes Matthias, Oesch Franz, Silva Renata, Ferreira Luísa M, Branco Paula S, Boer Douwe de, Remião Fernando, Carvalho Félix, Schön Michael R, Krebsfaenger Niels, Doehmer Johannes, Bastos Maria de Lourdes, Hengstler Jan G
REQUIMTE, Toxicology Department, Faculty of Pharmacy, University of Porto, Porto, Portugal.
Pharmacogenet Genomics. 2006 Nov;16(11):789-99. doi: 10.1097/01.fpc.0000230419.05221.fc.
Remarkable interindividual differences in 3,4-methylenedioxymethamphetamine ('Ecstasy')-mediated toxicity have been reported in humans. Therefore, we tested whether CYP2D6 or its variant alleles as well as CYP3A4 influence the susceptibility to 3,4-methylenedioxymethamphetamine.
3,4-Methylenedioxymethamphetamine cytotoxicity was determined in V79 cells expressing human wild-type CYP2D6 (CYP2D61), the low-activity alleles CYP2D62, *9, *10, and *17, as well as human CYP3A4. Metabolites of 3,4-methylenedioxymethamphetamine formed by the different cell lines were quantified by high-performance liquid chromatography/electrochemical detector.
Toxicity of 3,4-methylenedioxymethamphetamine was clearly increased in cells expressing CYP2D61 compared with the parental cells devoid of CYP-dependent enzymatic activity. Toxicity in V79 CYP2D61 cells was also higher than in V79 cell lines expressing the low-activity alleles CYP2D6*2, *9, *10, or 17. In contrast to CYP2D6, the CYP3A4 isoenzyme did not enhance 3,4-methylenedioxymethamphetamine toxicity. Formation of the oxidative 3,4-methylenedioxymethamphetamine metabolite N-methyl-alpha-methyldopamine was greatly enhanced in V79 cell line transfected with CYP2D61 compared to all other cell lines. The increase in the cytotoxic effects of 3,4-methylenedioxymethamphetamine observed in this cell line was therefore suspected to be a consequence of the production of this metabolite. This was further investigated by testing the cytotoxicity of N-methyl-alpha-methyldopamine to the control cell line. The results confirmed our hypothesis as the metabolite proved to be more than 100-fold more toxic than the parent compound 3,4-methylenedioxymethamphetamine.
CYP2D6*1 mediates 3,4-methylenedioxymethamphetamine toxicity via formation of N-methyl-alpha-methyldopamine. Therefore, it will be important to investigate whether CYP2D6 ultrarapid metabolizers are overrepresented in the cases of 3,4-methylenedioxymethamphetamine intoxications.
据报道，人类在3,4-亚甲基二氧甲基苯丙胺（“摇头丸”）介导的毒性方面存在显著的个体差异。因此，我们测试了CYP2D6及其变异等位基因以及CYP3A4是否会影响对3,4-亚甲基二氧甲基苯丙胺的易感性。
在表达人类野生型CYP2D6（CYP2D61）、低活性等位基因CYP2D62、*9、10和17以及人类CYP3A4的V79细胞中测定3,4-亚甲基二氧甲基苯丙胺的细胞毒性。通过高效液相色谱/电化学检测器对不同细胞系形成的3,4-亚甲基二氧甲基苯丙胺代谢物进行定量。
与缺乏CYP依赖性酶活性的亲本细胞相比，表达CYP2D61的细胞中3,4-亚甲基二氧甲基苯丙胺的毒性明显增加。V79 CYP2D61细胞中的毒性也高于表达低活性等位基因CYP2D6*2、9、10或17的V79细胞系。与CYP2D6不同，CYP3A4同工酶并未增强3,4-亚甲基二氧甲基苯丙胺的毒性。与所有其他细胞系相比，用CYP2D61转染的V79细胞系中氧化型3,4-亚甲基二氧甲基苯丙胺代谢物N-甲基-α-甲基多巴胺的形成大大增强。因此，怀疑该细胞系中观察到的3,4-亚甲基二氧甲基苯丙胺细胞毒性增加是这种代谢物产生的结果。通过测试N-甲基-α-甲基多巴胺对对照细胞系的细胞毒性对此进行了进一步研究。结果证实了我们的假设，因为该代谢物被证明比母体化合物
